Celllight golgi rfp

Manufactured by Thermo Fisher Scientific

Sourced in United States

The CellLight Golgi-RFP is a fluorescent protein-based reagent that specifically labels the Golgi apparatus in live cells. It is designed to enable real-time visualization and monitoring of the Golgi complex within the cellular environment.

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Lab products found in correlation

Hoechst 33342, by Thermo Fisher Scientific (6 mentions) Bacmam 2, by Thermo Fisher Scientific (4 mentions) Celllight er rfp, by Thermo Fisher Scientific (3 mentions) Bz 9000 fluorescence microscope, by Keyence (2 mentions) Dantrolene, by Bio-Techne (1 mentions) β nicotinamide adenine dinucleotide sodium salt, by Merck Group (1 mentions) Dantrolene sodium salt, by Bio-Techne (1 mentions) Er tracker blue white dpx, by Thermo Fisher Scientific (1 mentions) Cyclic adp ribose, by Merck Group (1 mentions) Celllight mitochondria gfp, by Thermo Fisher Scientific (1 mentions) Carmustine, by Merck Group (1 mentions) Paraformaldehyde (pfa), by Honeywell (1 mentions) Amplex red cholesterol assay kit, by Thermo Fisher Scientific (1 mentions) Immobilon p pvdf membrane, by Merck Group (1 mentions) Nunc lab tek 2 chambered coverglass, by Thermo Fisher Scientific (1 mentions) Immobilon western chemiluminescent hrp substrate, by Merck Group (1 mentions) Prosieve quadcolor protein marker, by Lonza (1 mentions) Turbofect transfection reagent, by Thermo Fisher Scientific (1 mentions) Pngase f, by New England Biolabs (1 mentions) Rc dc protein assay kit, by Bio-Rad (1 mentions)

13 protocols using celllight golgi rfp

Visualizing Golgi and Plasma Membrane

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ECV 304 (150,000 cells/well) or EA.hy 926 cells (250,000 cells/ well) were plated on 30 mm- or 1.5H high-precision- glass coverslips (Marienfeld-Superior, Lauda Königshofen, Germany) and grown until 70% confluency. Cells were infected with 30 particles per cell (ppc) of CellLight^® Golgi-RFP, BacMam 2.0 or 30 ppc of CellLight^® plasma membrane-RFP, BacMam 2.0 (ThermoFisher Scientific, Vienna, Austria) in culture medium containing antibiotics without serum. The cells were either imaged directly after l6h incubation with EV-HDL or EL-HDL or after a second round of infection.

Radulović S., Gottschalk B., Hörl G., Zardoya-Laguardia P., Schilcher I., Hallström S., Vujić N., Schmidt K., Trieb M., Graier W.F., Malli R., Kratky D., Marsche G, & Frank S. (2020). Endothelial lipase increases eNOS activating capacity of high-density lipoprotein. Biochimica et biophysica acta. Molecular and cell biology of lipids, 1865(4), 158612.

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Visualizing Organelle Dynamics in Living Cells

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β-Nicotinamide adenine dinucleotide sodium salt (Sigma Aldrich, N0632–1G), cyclic ADP ribose (Sigma Aldrich, C7344–5MG), Dantrolene sodium salt (Tocris, 0507), CellLight Mitochondria-GFP, BacMam 2.0 (Thermo Fisher Scientific, C10600), CellLight Golgi-RFP, BacMam 2.0 (Thermo Fisher Scientific, C10593), ER-Tracker Blue-White DPX (Thermo Fisher Scientific, E12353), Dantrolene (Tocris, 507). DPA/Terbium for membrane fusion assay kit (Biotium, 80104), 2,6-Pyridinedecarboxilic acid (Sigma Aldrich, P63808), Carmustine (Millipore Sigma, C0400).

Park J.H., Lo E.H, & Hayakawa K. (2021). Endoplasmic reticulum interaction supports energy production and redox homeostasis in mitochondria released from astrocytes. Translational stroke research, 12(6), 1045-1054.

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Biochemical Characterization of Proteins

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Unless otherwise specified, all chemicals and reagents, the ANTI-FLAG M2 affinity gel, the Immobilon-P PVDF Membrane (pore size: 0.45 μm) and Immobilon Western Chemiluminescent HRP Substrate were from Merck/Sigma-Aldrich and Millipore (Darmstadt, Germany). All cell culture media and supplements, the TurboFect transfection reagent, the Amplex^® Red Cholesterol assay kit, the CellLight™ Golgi-RFP, BacMam 2.0 Golgi-labelling reagent, and the eight-well microscopy plates Nunc™ Lab-Tek™ II Chambered Coverglass were from Thermo Fisher Scientific (Waltham, MA, USA). Paraformaldehyde was from Riedel-de Haën (Seelze, Germany), the RC-DC Protein Assay kit from BioRad (Hercules, CA, USA), the ProSieve^® QuadColor™ protein marker from Lonza (Basel, Switzerland) and the PNGase F was from New England Biolabs (Ipswich, MA, USA).

Dondapati D.T., Cingaram P.R., Ayaydin F., Nyeste A., Kanyó A., Welker E, & Fodor E. (2021). Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin. Membranes, 11(12), 978.

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Visualizing Golgi and Plasma Membrane

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Golgi Dynamics in A549 Cells

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A549 cells were transduced with CellLight Golgi‐RFP (Thermo Fischer Scientific, C10593) at a concentration of 40 particles per cell and cultured for 24 h. Subsequently, the cells were treated with vehicle or BFA (0.1 μg·mL⁻¹) for 6 h, and then, nuclei were stained with PureBlu Hoechst 33342 (Bio‐Rad, Hercules, CA, USA, 135‐1304). After fixation in 4% paraformaldehyde, fluorescence images were acquired using a BZ‐9000 fluorescence microscope (KEYENCE, Osaka, Japan).

Matsuo Y, & Hirota K. (2017). Transmembrane thioredoxin‐related protein TMX1 is reversibly oxidized in response to protein accumulation in the endoplasmic reticulum. FEBS Open Bio, 7(11), 1768-1777.

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Mitochondrial, Golgi, and ER Imaging

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SiR-CA was synthesized according to previously reported methods. BacMam 2.0 reagent, CellLight Mitochondria-GFP (PDHA1–GFP; C10600), CellLight Mitochondria-RFP (PDHA1–RFP; C10505), CellLight Golgi-GFP (GALNT1–GFP; C10592), CellLight Golgi-RFP (GALNT1–RFP; C10593), CellLight ER-GFP (KDEL–GFP; C10590), CellLight ER-RFP (KDEL–RFP; C10591), culturing medium DMEM (12430054, 21063029 and A1443001), Opti-MEM with no phenol red (11058021) and FBS and chemical reagents Hoechst 33342 (62249) and MitoTracker Deep Red FM (M22426) were all purchased from Thermo Fisher. CellTiter-Glo 2.0 (G9242) was purchased from Promega, and alexidine dihydrochloride (A8986) was purchased from Sigma-Aldrich. Plasmid encoding mEmerald–TOMM20 was a gift from M. Davidson (Florida State University, mEmerald–TOMM20-N-10, Addgene plasmid 54282). Plasmid encoding Halo-TOMM20 was a gift from K. McGowan (HHMI Janelia, Halo-TOMM20-N-10, Addgene plasmid 123284). Plasmid encoding COX8A-SNAP was a gift from A. Egana (New England Biolabs, pSNAPf-COX8A, Addgene plasmid 101129).

Zheng S., Dadina N., Mozumdar D., Lesiak L., Martinez K.N., Miller E.W, & Schepartz A. (2023). Long-term super-resolution inner mitochondrial membrane imaging with a lipid probe. Nature Chemical Biology, 20(1), 83-92.

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Visualizing CALN1 expression in HAC15 cells

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pLVSIN-N-AcGFP with CALN1 was transduced in HAC15 cells. The ER and Golgi bodies were fluorescently labeled using CellLight ER-RFP (Thermo Fisher Scientific, Waltham, MA, USA) and CellLight Golgi-RFP (Thermo Fisher Scientific, Waltham, MA, USA), respectively, following the manufacturer's protocol. The fluorescence was detected using the BZ-9000 fluorescence microscope (KEYENCE, Itasca, IL, USA).
To investigate endogenous CALN1 expression, HAC15 cells were first incubated with CellLight ER-RFP for 24 hours. The cells were then fixed for 10 minutes in 4% paraformaldehyde, and 0.5% Triton X-100 was added to the fixed cells for 10 minutes. The primary antibody for CALN1 (Abcam, #ab117584, Cambridge, UK) and ProLong Gold Antifade Reagent with DAPI (Thermo Fisher Scientific, Waltham, MA, USA) were then applied.

Kobuke K., Oki K., Gomez-Sanchez C.E., Gomez-Sanchez E.P., Ohno H., Itcho K., Yoshii Y., Yoneda M, & Hattori N. (2017). Calneuron 1 Increased Ca2+ in the Endoplasmic Reticulum and Aldosterone Production in Aldosterone-producing Adenoma. Hypertension (Dallas, Tex. : 1979), 71(1), 125-133.

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Golgi Apparatus Visualization in MDA-MB-231 Cells

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The glass bottom of the confocal dish was coated with fibronectin (FN). A total of 2 × 10⁵ cells were seeded and allowed to attach for 12 h before staining. The reagent CellLight Golgi-RFP (C10593, Thermofisher, Waltham, MA, USA) was added directly to the MDA-MB-231 cells. After incubated overnight, the cells were ready to image the next morning. Hoechst 33342 (H3570, Thermofisher, Waltham, MA, USA), diluted at 1:5000 in PBS, was added and incubated for at least 10 min. Cells were then washed twice, for 3 min each, in prewarmed PBS.

Guan L.Y., Lv J.Q., Zhang D.Q, & Li B. (2021). Collective Polarization of Cancer Cells at the Monolayer Boundary. Micromachines, 12(2), 112.

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Multidrug Combination Therapy Evaluation

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Doxorubicin (Dox, MB1087-1), Methotrexate (MTX, MB1156-1), Cisplatin (Cis, MB1055-1) and Paclitaxel (PTX, MB1178-1) were purchased from Meilunbio. Amiloride hydrochloride (PHR1839), NAC (A9165), DPI (43088), cyclosporin A (CsA, SML1018), EHT1864 (E1657), SKF-525A (567300), SB203580 (S8307), JSH-23 (J4455), PKH26-Red fluorescent cell linker kit, NH₄Cl (A9434), PKH67-Green fluorescent cell linker kit and anti-Lamp1 antibody (L1418) were purchased from Sigma-Aldrich. Ryanodine (ab120083), CGP37157 (ab120012) were purchased from Abcam. Bafilomycin A1 (HY-100558) were purchased from MCE. ER-, Mito-, Lyso-Tracker Green/Red, LysoSensor Yellow/Blue DND-160 (L7545), Early Endosomes-GFP (C10586), LysoSensor Green DND-189 (L7535), CellLight Golgi-RFP (C10593) and Late Endosomes-GFP (C10588) were purchased from Thermo Fisher Scientific. Clodronate liposomes were purchased from Liposomal.

Wei K., Zhang H., Yang S., Cui Y., Zhang B., Liu J., Tang L., Tan Y., Liu S., Chen S., Yuan W., Luo X., Chen C., Li F., Liu J., Chen J., Xu P., Lv J., Tang K., Zhang Y., Ma J, & Huang B. (2023). Chemo-drugs in cell microparticles reset antitumor activity of macrophages by activating lysosomal P450 and nuclear hnRNPA2B1. Signal Transduction and Targeted Therapy, 8, 22.

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Investigating STING and IRF3 Signaling

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STING (Cat# D2P2F), phospho-STING S366 (Cat# D7C36), IRF3 (Cat# D83B9),
and phospho-IRF3 S396 (Cat# D601M) were purchased from Cell Signaling
Technology; DNA (Cat# CBL186; 1:250) from Thermo Fisher; VDAC1 (Cat#
NB100–695) from Novus Biologicals; cGAS (Cat# sc-515777), TOM20 (Cat#
sc-11415), and GAPDH (Cat# sc-47724) from Santacruz Biotechnology; and
α-smooth muscle actin (Cat# MAB1420; 1:500) from R&D systems. Human
recombinant TGF-β was purchased from R&D systems (7754-BH). Inhibitor
for STING (Cat# IFM-004490–7) was provided by IFM therapeutics. The CGAS
inhibitor RU.521 (Cat# SML2347–5MG), 4-Dinitrophenol (Cat# D-198501),
SMAD inhibitor (Cat# SB53512), and zinc chelator N, N, N′,
N′-Tetrakis (2-pyridylmethyl) ethylenediamine TPEN (Cat# P-4413) were
purchased from Millipore Sigma. Golgi were labeled using CellLight Golgi-RFP
(Thermo Fisher Scientific, Cat# C10593). The CellLight reagent was directly
added to medium (2 μL/200 μL medium) before treatment. Picogreen
stain was from the Quant-IT assay kit (Thermo Fisher Scientific, Cat#
P7589).

Arumugam S., Li B., Boodapati S.L., Nathanson M.H., Sun B., Ouyang X, & Mehal W.Z. (2023). Mitochondrial DNA and the STING pathway are required for hepatic stellate cell activation. Hepatology (Baltimore, Md.), 78(5), 1448-1461.

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